Authors:Hong-Tae Park; Hyun-Eui Park; Young-Hoon Jung; Han Sang YooPages: 1 - 6Abstract: Publication date: March 2018 Source:Veterinary Microbiology, Volume 216 Author(s): Hong-Tae Park, Hyun-Eui Park, Young-Hoon Jung, Han Sang Yoo Mycobacterium avium subsp. paratuberculosis (MAP) is a causative agent of Johne’s disease or paratuberculosis (PTB), which is a chronic debilitating disease in ruminants, that is characterized by incurable enteritis and persistent diarrhea. ISMap02 is one of the major targets of PCR because it is present in multicopies (six copies) and known to be specific to MAP. However, in the present study, non-MAP mycobacteria were shown to be positive by ISMap02 targeting PCR. Two bacterial isolates (Sample ID: BO-038 and BO-042) were cultured from bovine fecal samples that produced positive results in three of two ISMap02 targeting PCR analyses with negative results in IS900 real-time PCR. Species identification using 16S rRNA gene sequencing and hsp65 gene partial sequencing revealed that strains BO-038 and BO-042 were M. virginiense and M. nonchromogenicum, respectively, which both belong to the M. terrae complex (MTC). Moreover, the two isolates shared a novel insertion sequence (IS) with high similarity to some parts of nucleotide sequences of ISMap02, and IS was presumed to be identical to that present in M. heraklionense. Both the novel IS and ISMap02 were characterized as IS1182 family members, and several sequences similar to ISMap02 were identified by BLAST analysis. In addition, the DDE transposase of the novel IS showed great similarity in the N-terminal portion with the IS5/1182 DDE transposase of other mycobacteria. These results suggest that ISMap02 has a conserved region with similarity to other ISs, and that the diagnostic value of the primer sets targeting that region should be re-addressed.

Authors:Yu Jia-yu; Zhu Qian; Diao Fei-fei; Teng Chuan-jie; Peng Hui; Shang Yuan-yuan; Zhao Yong-feng; Wang Jian-li; Shijin Jiang; Xie Zhi-jingPages: 7 - 12Abstract: Publication date: March 2018 Source:Veterinary Microbiology, Volume 216 Author(s): Yu Jia-yu, Zhu Qian, Diao Fei-fei, Teng Chuan-jie, Peng Hui, Shang Yuan-yuan, Zhao Yong-feng, Wang Jian-li, Shijin Jiang, Xie Zhi-jing Three parvoviruses were isolated from the raccoon dogs experiencing severe enteritis, named RDPV-DP1, RDPV-DP2 and RDPV-DP3, respectively. The VP2 genes of the 3 isolates showed 99.9% identity at the nucleotide level, and shared 99.1%-99.5% identity with the reference CPVs. The RDPVs resembled original CPV-2, but with four mutations. The RDPVs displayed S297A of VP2 protein as CPV-2a or CPV-2b prevalent throughout most of the world. Residue N375D was found in the 3 isolates, resembling CPV-2a/2b/2c. And the 3 isolates had a natural mutation of VP2 residue V562L, which is adjacent to residue 564 and 568 and might be involved in host range. Interestingly, VP2 S27T was firstly found in the isolates. Phylogenetic analysis of VP2 genes revealed that the RDPVs were clustered into one small evolutionary branch and shared the identical branch with 7 CPV-2 isolates from raccoon dogs and one CPV-2 isolate from fox, not with CPV vaccine viruses. Phylogenetic analysis of NS1 genes demonstrated that the RDPVs shared the identical branch with the reference CPV-2a/2b/2c. Experimental infection showed that RDPV infection caused a high morbidity in raccoon dogs. It implied that the RDPV was virulent to raccoon dogs and continued to evolve in China.

Authors:Sandrine Baron; Laetitia Le Devendec; Fabrice Touzain; Eric Jouy; Pierrick Lucas; Claire de Boisséson; Emeline Larvor; Isabelle KempfPages: 20 - 24Abstract: Publication date: March 2018 Source:Veterinary Microbiology, Volume 216 Author(s): Sandrine Baron, Laetitia Le Devendec, Fabrice Touzain, Eric Jouy, Pierrick Lucas, Claire de Boisséson, Emeline Larvor, Isabelle Kempf Resistance to extended-spectrum cephalosporins (ESCs) is mostly borne by conjugative plasmids. The aim of the present study was to evaluate the characteristics and diversity of ESC resistance plasmids in Escherichia coli from different free-range broiler flocks in France, and their persistence in flocks during rearing. Two hatcheries were selected. Faecal samples from 11 flocks were collected from before their arrival on the broiler production farm up to their slaughter at the end of the rearing period. A selection of 25 E. coli isolates obtained at different times from different flocks but all harbouring an ESC resistance gene was characterised. The plasmids coding for ESC resistance were sequenced using Mi-seq Illumina technology or the ion proton system (Ion Torrent). Ten IncI1 ST12 plasmids carried the bla CMY-2 gene, and most of them had no other resistance genes. All bla CMY-2 plasmids were obtained from day-old to 7-day-old chicks from four flocks hatched at the same hatchery and sent to three different farms. Sequence comparisons showed identity percentages higher than 99%. Fifteen IncI1 ST3 plasmids were obtained from day-old to 77-day-old broilers from seven flocks on six farms. These plasmids harboured the bla CTX-M-1 gene, and most also had the tet(A) and sul2 genes, with sequence identity higher than 99%. For both types of plasmid, very high identity percentages were also obtained with published sequences of plasmids isolated from broilers in other countries or from other animal species. Thus, unlike the IncI1 ST12 bla CMY-2 plasmids, the epidemic nature of the IncI1 ST3 bla CTX-M-1 plasmids in the French poultry production makes it difficult to determine the origin of a contamination which may persist for weeks in a flock.

Authors:Agnieszka Sałamaszyńska-Guz; Ilona Stefańska; Paweł Bącal; Marian BinekPages: 25 - 30Abstract: Publication date: March 2018 Source:Veterinary Microbiology, Volume 216 Author(s): Agnieszka Sałamaszyńska-Guz, Ilona Stefańska, Paweł Bącal, Marian Binek A total of 43 Campylobacter isolates from poultry, cattle and pigs were investigated for their ability to form biofilm. The studied strains were also screened for motility, adhesion and invasion of Caco-2 cells as well as extracellular DNase activity. The relation between biofilm formation and selected phenotypes was examined. Biofilm formation by the tested strains was found as irrespective from their motility and not associated with colonization abilities of human Caco-2 cells. Results of our study show that Campylobacter isolates from various animal sources are able to form biofilm and invade human Caco-2 cells in vitro.

Authors:Áine B. Collins; John F. Mee; Peter D. KirklandPages: 31 - 37Abstract: Publication date: March 2018 Source:Veterinary Microbiology, Volume 216 Author(s): Áine B. Collins, John F. Mee, Peter D. Kirkland Schmallenberg virus (SBV) and Akabane virus (AKAV) are teratogenic Simbu serogroup Orthobunyaviruses. Embryonated chicken egg models (ECE) have been used to study the pathogenicity and teratogenicity of Simbu viruses previously, however to date no such studies have been reported for SBV. Hence, the aims of this study were to investigate if ECE are susceptible to experimental SBV infection, and to evaluate the pathogenicity and teratogenicity of SBV and AKAV in ECE models. Two studies were conducted. In Study A, SBV (106.4 TCID50) was inoculated into the yolk-sac of 6-day-old and 8-day-old ECEs. In Study B, SBV and AKAV were inoculated into 7-day-old ECEs at a range of doses (102.0–106.0 TCID50). ECE were incubated at 37 °C until day 19, when they were submitted for pathological and virological examination. SBV infection in ECE at 6, 7 and 8 days of incubation resulted in stunted growth and musculoskeletal malformations (arthrogryposis, skeletal muscle atrophy, contracted toes, distorted and twisted legs). Mortality was greater in embryos inoculated with SBV (31%) compared to AKAV (19%), (P < 0.01), suggesting that SBV was more embryo-lethal. However, embryos infected with AKAV had a significantly higher prevalence of stunted growth (P < 0.05) and musculoskeletal malformations (P < 0.01), suggesting that AKAV was more teratogenic in this model. These studies demonstrate for the first time that the ECE model is a suitable in vivo small animal model to study SBV. Furthermore, these results are consistent with the clinico-pathological findings of natural SBV and AKAV infection in ruminants.

Authors:Priscila Regina Guerra; Ana Herrero-Fresno; Susanne Elisabeth Pors; Shahana Ahmed; Dan Wang; Ida Thøfner; Fabio Antenucci; John Elmerdahl OlsenPages: 38 - 44Abstract: Publication date: March 2018 Source:Veterinary Microbiology, Volume 216 Author(s): Priscila Regina Guerra, Ana Herrero-Fresno, Susanne Elisabeth Pors, Shahana Ahmed, Dan Wang, Ida Thøfner, Fabio Antenucci, John Elmerdahl Olsen Over the last few years, polyamines have been described as key-signal of virulence in pathogenic bacteria. In the current study, we investigated whether the knockout of genes related to polyamine biosynthesis and putrescine transport affected the virulence of an avian pathogenic E. coli (APEC) strain. One-week-old White Leghorn chickens were infected intratracheally with mutants in polyamine biosynthesis (ΔspeB/C and ΔspeD/E) and transport genes (ΔpotE) of a well-characterized APEC strain of ST117 (O83: H4). All polyamine mutants and the wild-type strain were able to infect chicken; however, we observed significantly fewer lesions in the lungs of the chickens infected with the polyamine mutants in comparison with chicken infected with the wild-type. Results derived from histology of infected lungs detected significantly fewer lesions in the lung of birds infected within particular the putrescine transport mutant (ΔpotE). A decrease in colonization levels was observed in the liver and spleen of birds infected with the putrescine biosynthesis mutant ΔspeB/C, and likewise, a decrease of the colonization levels of all organs from birds infected with the ΔpotE was detected. Together, our data demonstrate that the deletion of polyamine genes, and in particular the PotE membrane protein, attenuates the virulence of APEC during infection of chickens.

Authors:Vera Haapala; Tarja Pohjanvirta; Nella Vähänikkilä; Jani Halkilahti; Henri Simonen; Sinikka Pelkonen; Timo Soveri; Heli Simojoki; Tiina AutioPages: 60 - 66Abstract: Publication date: March 2018 Source:Veterinary Microbiology, Volume 216 Author(s): Vera Haapala, Tarja Pohjanvirta, Nella Vähänikkilä, Jani Halkilahti, Henri Simonen, Sinikka Pelkonen, Timo Soveri, Heli Simojoki, Tiina Autio Mycoplasma bovis infections are responsible for substantial economic losses in the cattle industry, have significant welfare effects and increase antibiotic use. The pathogen is often introduced into naive herds through healthy carrier animals. In countries with a low prevalence of M. bovis, transmission from less common sources can be better explored as the pathogen has limited circulation compared to high prevalence populations. In this study, we describe how M. bovis was introduced into two closed and adequately biosecure dairy herds through the use of contaminated semen during artificial insemination (AI), leading to mastitis outbreak in both herds. Epidemiological analysis did not reveal an infection source other than semen. In both farms the primary clinical cases were M. bovis mastitis in cows inseminated with the semen of the same bull four weeks before the onset of the disease. One semen straw derived from the semen tank on the farm and other semen lots of this bull were positive for M. bovis. In contrast, semen samples were negative from other bulls that had been used for insemination in previous or later oestrus to those cows with M. bovis mastitis. Furthermore, cgMLST of M. bovis isolates supported the epidemiological results. To our knowledge this is the first study describing the introduction of M. bovis infection into a naive dairy herd via processed semen. The antibiotics used in semen extenders should be re-evaluated in order to provide farms with M. bovis-free semen or tested M. bovis-free semen should be available.

Authors:Xinxin Niu; Lanlan Liu; Chunyan Han; Jing Li; Xiangwei ZengPages: 67 - 71Abstract: Publication date: Available online 6 February 2018 Source:Veterinary Microbiology Author(s): Xinxin Niu, Lanlan Liu, Chunyan Han, Jing Li, Xiangwei Zeng To probe the epidemiology of duck circovirus (DuCV) in migrating wild ducks in China, 189 samples collected from 11 species of wild ducks from 2013 to 2016 were analyzed by PCR. Four positive samples were obtained from Mallard (2), Green-winged Teal (1), and Falcated Duck (1), and the positive rate was 2.12%. The homologous alignment of the complete genome shows that the homology of the wild duck strains is 81.6 to 95.1% to each other and 81.4 to 100% to the 32 strains from other origins. Amino acids alignment revealed that the mutable ORF2 gene has six major variable regions and many random mutation points. Phylogenetic analysis showed that the four sequences belong to two genotypes: wd2013017, wd2013046 and AY228555 (representative strain of genotype 1) belong to genotype 1, while wd2014012, wd2015028 and AY394721 (representative strain of genotype 2) belong to genotype 2. The results indicate that both genotypes of DuCV are distributed and common in migrating wild ducks. Further analysis shows that duck circovirus infection is mainly concentrated in the eastern coastal cities of China, which are part of the East Asian-Australian flyway. This suggests that wild ducks with circovirus may be an important factor in the epidemic and spread of DuCV.

Authors:Luca Ferrari; Elena Canelli; Elena De Angelis; Alessia Catella; Giulia Ferrarini; Giulia Ogno; Luca Bonati; Roberto Nardini; Paolo Borghetti; Paolo MartelliPages: 85 - 92Abstract: Publication date: Available online 6 February 2018 Source:Veterinary Microbiology Author(s): Luca Ferrari, Elena Canelli, Elena De Angelis, Alessia Catella, Giulia Ferrarini, Giulia Ogno, Luca Bonati, Roberto Nardini, Paolo Borghetti, Paolo Martelli Highly pathogenic (HP) PRRSV isolates have been discovered within both PRRSV-1 and PRRSV-2 genotypes and investigated in recent years especially for their ability to cause extremely severe disease in conventional pig herds. The exacerbation of general and respiratory clinical signs has been attributed not only to an efficient replication (virulence) but also to the ability to dysregulate viral recognition and induce mechanisms of immune evasion or immune enhancement of humoral and cellular anti-viral responses differently from non-HP PRRSV isolates in terms of intensity and temporal onset. Thus, the understanding of the immunopathogenesis of HP PRRSV is a major concern for the study of virus biology and development of efficacious vaccines. The present study aims at addressing the modulation of relevant immune cell subsets by flow cytometry in the blood of 4-week-old pigs experimentally infected with the recently discovered PR40/2014 HP PRRSV-1.1 strain phenotypically characterized in Canelli et al. (2017) compared to pigs infected with a non-HP PRRSV isolate (PR11/2014) and uninfected controls. PR40 infected animals showed an early and marked reduction of pro-inflammatory CD172α+ CD14+CD16+ and CD14+CD163+ monocytes and TCRγδ+ CD8α + /CD8α- lymphocytes when pigs were most infected, possibly due to a recruitment sustaining an acute inflammatory response in target tissues. The prolonged increased CD3+CD16+ NKT cell levels may sustain peripheral inflammation and/or the anti-viral response. The late reduction (potential depletion) of γ/δ T lymphocytes and CD3+CD4+CD8α- naïve Th lymphocytes paralleled with the delayed increase of CD3+CD4+CD8α+ memory and CD3+CD4-CD8α/β+ cytotoxic T lymphocytes. In addition, PR40 infection showed an early depletion of activated CD4+CD25+ T lymphocytes and Tregs together with an intense and lasting depletion of CD21+ B lymphocytes. Overall, these features demonstrate that the more severe clinical signs observed upon infection with the HP PR40 strain are sustained by remarkable changes in the peripheral blood distribution of immune cells and provide further insights into the immune regulation/immunopathogenesis induced by PRRSV-1 subtype 1 European isolates.

Authors:David A. Jaffe; Bruno B. Chomel; Rickie W. Kasten; Edward B. Breitschwerdt; Ricardo G. Maggi; Ashleigh McLeish; Ulrike ZiegerPages: 119 - 122Abstract: Publication date: Available online 9 February 2018 Source:Veterinary Microbiology Author(s): David A. Jaffe, Bruno B. Chomel, Rickie W. Kasten, Edward B. Breitschwerdt, Ricardo G. Maggi, Ashleigh McLeish, Ulrike Zieger Many mammals are established hosts for the vector borne bacterial genus, Bartonella. Small Indian mongooses (Herpestes auropunctatus) have only been reported as a possible host for Bartonella henselae in southern Japan. Confirming Bartonella presence in mongooses from other regions in the world may support their role as potential reservoirs of this human pathogen. Specifically, documenting Bartonella in Caribbean mongooses would identify a potential source of zoonotic risk with mongoose-human contact in the New World. Using serological and molecular techniques, we investigated B. henselae DNA and specific antibody prevalence in 171 mongooses from all six parishes in Grenada, West Indies. Almost a third (32.3%, 54/167) of the tested mongooses were B. henselae seropositive and extracted DNA from 18/51 (35.3%) blood pellets (n=51) were PCR positive for the citrate synthase (gltA) and/or the β subunit of RNA polymerase (rpoB) genes. All sequences were identical to B. henselae genotype I, as previously reported from Japan. This study confirms the role of small Indian mongooses as a natural reservoir of B. henselae in the New World.

Authors:Haijin Liu; Renata Servan de Almeida; Patricia Gil; Emmanuel AlbinaPages: 123 - 131Abstract: Publication date: Available online 9 February 2018 Source:Veterinary Microbiology Author(s): Haijin Liu, Renata Servan de Almeida, Patricia Gil, Emmanuel Albina Newcastle disease, caused by the virulent Newcastle disease virus (NDV), poses a risk for the poultry industry. The virulence of NDV is mainly determined by the cleavage site of F protein. Lentogenic NDV can become velogenic after passages in chicken brain and air sac, because the proportion of virulent-related strains gradually increases. In contrast, this proportion remains unchanged if NDV is passaged via chicken embryos. This information suggests that environmental conditions rather than mutation affect NDV fitness in quasispecies. However, it is unknown how the environment selects virulent-related strains from a viral population. In this study, velogenic and lentogenic NDV marked by green or red fluorescence were used to establish persistent infection (PI) in BHK-21 cells. Monitoring viruses by different methods, we found that, without competition, persistently infected cells harbored lentogenic and velogenic NDV strains similarly in terms of viral release, viral spread and the period of persistent viral infection. In contrast, under competitive co-infection, velogenic NDV became dominant in quasispecies from the fifth passage of PI cells, which resulted in the progressive disappearance of the lentogenic NDV strain. This domination was concomitant with a short-term reduction in the superinfection exclusion and supernatant interference in PI cells resulting in a velogenic virus rebound. We concluded that virulent-related F protein cleavage site facilitates the spread and replication of NDV in conditions under which cells do not secret trypsin-like proteases and do not inhibit free virus infection, resulting in a gradual increase in virulent strains in quasispecies with the number of passages.

Authors:Raquel Portugal; Alexandre Leitão; Carlos MartinsPages: 132 - 141Abstract: Publication date: Available online 7 February 2018 Source:Veterinary Microbiology Author(s): Raquel Portugal, Alexandre Leitão, Carlos Martins ASFV causes an important disease of domestic swine and wild boar. Currently no vaccine is available, highlighting the necessity to understand ASFV modulation of innate immune responses in natural host cells. With this aim, macrophage cultures enriched in SWC9 and CD163 differentiation markers were infected in parallel with high virulent ASFV/L60 and low virulent ASFV/NHV, the latter lacking MGF 360 and 505/530 genes associated with type I interferon (IFN I) control. IFN I production and signaling were studied after completion of the viral cycles. None of the viruses increased IFN I production in host cells, and accordingly, didn't cause activation of the central mediator of the pathway IRF3. However, upon stimulation by poly:IC treatment during infections, L60 and NHV similarly inhibited IFN I production. This didn't seem to depend on IRF3 modulation since its activation levels were not significantly decreased in L60 infection and were even increased in NHV's, in comparison to stimulated mock infections. The infections didn't evidently activate JAK-STAT pathway mediators STAT1 and STAT2, but did increase expression of interferon stimulated genes (ISGs), to higher levels in NHV than L60 infection. Interestingly, in presence of IFN-α, L60 but not NHV was able to decrease significantly the expression of some of the ISGs tested. Overall, both L60 and NHV were able to inhibit IFN I production in macrophages, through a mechanism not dependent on IRF3 modulation. The high virulent isolate showed however a more effective control of the downstream ISGs expression pathway.

Authors:Deepti K. Pillai; Elva Cha; Derek MosierPages: 11 - 17Abstract: Publication date: February 2018 Source:Veterinary Microbiology, Volume 215 Author(s): Deepti K. Pillai, Elva Cha, Derek Mosier Bovine respiratory disease (BRD) is a major problem for the cattle industry that is triggered by various environmental stressors, pathogens and host responses. Mannheimia hemolytica, an important bacterial component of BRD, are present within the nasopharayngeal region of normal calves as commensal biofilm communities. However, following stress there are changes in the nasopharyngeal microenvironment that triggers the transition of the commensal M. haemolytica into a pulmonary pathogen. The factors responsible for this transition in- vivo are unknown. In this study we developed an in-vitro biofilm model and investigated the effect of three stress- related compounds: norepinephrine (NE), epinephrine (E), and substance P (SP) on M. haemolytica biofilms. Biofilm formation was demonstrated for 3 bovine nasal isolates of M. haemolytica by growing them in basal culture media, basal media with additional glucose, and basal media with a reduced pH. Increased glucose enhanced biofilm biomass for 2/3 isolates, but acidic media did not increase biofilm biomass when compared to biofilm biomass in basal media. When the biofilm was exposed to NE, E and SP, there was a dispersal of the biofilm which was most effective with E, followed by NE, and SP being the least effective. Using high – throughput scanning electron microscopy and confocal-imaging we confirmed our experimental data that treatment with NE, E and SP cause dispersion of M.haemolytica from biofilms.

Authors:Cecilia Cundon; Claudia Carolina Carbonari; Gisela Zolezzi; Marta Rivas; Adriana BentancorPages: 29 - 34Abstract: Publication date: February 2018 Source:Veterinary Microbiology, Volume 215 Author(s): Cecilia Cundon, Claudia Carolina Carbonari, Gisela Zolezzi, Marta Rivas, Adriana Bentancor Shiga toxin-producing Escherichia coli (STEC) is th etiological agent of gastrointestinal diseases as haemorrhagic colitis and haemolytic-uraemic syndrome (HUS). Shiga toxin (Stx) is the main virulence factor. There are two types, Stx1 and Stx2, and several subtypes. Other virulence factors are involved in pathogenesis. While O157:H7 is the predominant serotype, at present non-O157 STEC strains are increasingly recognized as foodborne pathogens worldwide. In Argentina, STEC O174 stands out as an emerging pathogen and is one of the four most prevalent non-O157 STEC serogroups. The aim of this study was to characterize 41 O174:[H21, H28] STEC strains isolated from animals, food, and humans. Isolates were characterized by stx genotyping, adhesion factors (afaC, eae, iha, lpf O113, saa, and toxB), additional toxins (cdtV, ehxA, subA) and clonal relationship by pulsed-field gel electrophoresis (PFGE). Among 30 O174:H21 strains, the most prevalent stx subtype was stx 2c (56.7%), and among 11 O174:H28 strains, the most prevalent was stx 2a (90.9%). Regarding virulence factors, all strains were positive for afaC gene and negative for eae and toxB genes. In O174:H21, the frequency of additional factors was lpf O113 (96.6%), iha (83.3%), ehxA and saa (10%), and subA (3.3%), meanwhile in O174:H28 strains the frequency was iha and subA (100%), lpf O113 (90.9%), ehxA and saa (90.9%), and cdtV (9.09%). By Xbal-PFGE, 29 patterns were established with 64.01% similarity and three clusters were detected. Given the fact that the O174 serogroup is a local emergent, it is important to study the virulence profiles to understand its potential pathogenicity.

Authors:Michał. K. Krzysiak; Artur Jabłoński; Wojciech Iwaniak; Monika Krajewska; Julia Kęsik-Maliszewska; Magdalena LarskaPages: 57 - 65Abstract: Publication date: February 2018 Source:Veterinary Microbiology, Volume 215 Author(s): Michał. K. Krzysiak, Artur Jabłoński, Wojciech Iwaniak, Monika Krajewska, Julia Kęsik-Maliszewska, Magdalena Larska After the complete extinction from the wild of European bison (Bison bonasus) at the beginning of the twentieth century, the worldwide species population was restored to approximately 5500 individuals, with the species however remaining endangered. Despite numerous studies on the ecology and genetics of European bison, the threats of infectious diseases have been largely unexamined. The aim of this study was to screen the exposure of the world’s largest population of European bison to the pathogens, which may influence the condition and development of the endangered species. A total of 240 free-ranging and captive European bison from eight main Polish populations sampled were tested for the presence of specific antibodies against ten different viruses, bacteria or protozoan. The samples were collected from chemically immobilized, selectively culled or found dead animals. Based on serology, the exposure to bovine viral diarrhea virus (BVDV), bovine herpesvirus type 1 (BoHV-1), Mycoplasma and Brucella spp. was determined as rather accidental. Using gamma-interferon assay followed by Mycobacterium tuberculosis subs. caprae detection in tissues, diagnosis of bovine tuberculosis was made for 6 out of 78 (7.7%) bison from one captive herd. The highest seroprevalence was found for bovine adenovirus type 3 (BAdV-3) −60.2% and bovine parainfluenza type 3 (PIV-3) −34.0%, while the antibodies against bovine respiratory syncytial virus (BRSV), Toxoplasma gondii and Leptospira spp. were found in 10.4%, 10.4% and 8.7% of samples, respectively. In the multivariable statistical analysis using generalized linear mixed models (GLMMS), the risk factors for PIV-3 seropositivity included population type (free-living/captive), age and health status (apparently healthy/eliminated due to the poor condition). Higher risk of BAdV-3 seropositive result was observed in free-living female European bison. The high BAdV-3 and PIV-3 seroprevalences may suggest involvement of these pathogens in the most frequently observed respiratory disorders in European bison. Moreover, this is the first study demonstrating BAdV-3 exposure in the species.

Authors:B. Schauer; R. Krametter-Frötscher; F. Knauer; R. Ehricht; S. Monecke; A.T. Feßler; S. Schwarz; T. Grunert; J. Spergser; I. LoncaricPages: 77 - 82Abstract: Publication date: February 2018 Source:Veterinary Microbiology, Volume 215 Author(s): B. Schauer, R. Krametter-Frötscher, F. Knauer, R. Ehricht, S. Monecke, A.T. Feßler, S. Schwarz, T. Grunert, J. Spergser, I. Loncaric The aim of this study was to determine the prevalence, the antimicrobial resistance patterns and the genetic diversity of methicillin-resistant Staphylococcus aureus (MRSA) from Austrian ruminants and New World camelids that were treated at the University of Veterinary Medicine, Vienna. Between April 2014 and January 2017, 723 nasal swabs originating from ruminants and New World camelids were examined. MRSA isolates were characterized by mecA/mecA1/mecC PCRs and by DNA microarray analysis. They were genotyped by spa typing, dru typing, MLST and MLVA. Glycopolymer fingerprinting by FTIR spectroscopy was also performed. Antimicrobial susceptibility testing was conducted by agar disk diffusion. Twelve MRSA isolates were mecA-positive, whereas three were mecC-positive. The MRSA isolates carried five different SCCmec elements, and belonged to three sequence types (ST45, ST130, ST398). The MRSA isolates displayed seven different resistance phenotypes. The present study describes for the first time mecC-carrying MRSA isolates originating from domesticated animals in Austria. More systematic studies are needed to unravel the role of ruminants and New World camelids as reservoirs for MRSA as a potential risk for zooanthropogenic transmission.

Authors:K.K. Patel; L. Howe; N. Haack; C. Heuer; G.W. Asher; P.R. WilsonPages: 83 - 89Abstract: Publication date: February 2018 Source:Veterinary Microbiology, Volume 215 Author(s): K.K. Patel, L. Howe, N. Haack, C. Heuer, G.W. Asher, P.R. Wilson This paper investigates Leptospira borgpeterseni serovar Hardjobovis and L. interrogans serovar Pomona as potential causes of sub-optimum pregnancy rates and mid-term abortion in farmed red deer. Rising two-year-old (R2, n = 22,130) and mixed-age (MA, n = 36,223) hinds from 87 and 71 herds, respectively, throughout New Zealand were ultrasound scanned early in gestation (Scan-1) and a sub-sample re-scanned (Scan-2) 55–89 days later and mid-term daily abortion rate calculated. A sub-sample of sera from pregnant and non-pregnant hinds at both scans, both with (case herds) and without aborted hinds was tested for Leptospira using the microscopic agglutination test with titre cut-point ≥1:48 as positive. At Scan-1, 44.3% of 661 and 4.6% of 647 hinds were sero-positive for Hardjobovis and Pomona, respectively. The geometric mean titre (GMT) for Pomona was higher in pregnant than non-pregnant MA hinds (p = 0.015) but not in R2 hinds. At Scan-2 in case herds, 40.3% of 2242 and 7.1% of 2243 hinds were sero-positive for Hardjobovis and Pomona, respectively. There was no association between Hardjobovis or Pomona serology and non-pregnancy (Scan-1) or mid-term abortion (Scan-2) at animal or herd level. In case herds, GMT was higher in non-aborted than aborted hinds for Hardjobovis (p = 0.018) in MA herds and for Pomona in R2 herds (p = 0.006). No uteri from hinds not pregnant or aborting at either scan, or not rearing a calf to weaning, and fetuses as available, were positive on PCR. Evidence is insufficient to confirm that Leptospira Hardjobovis and Pomona play a significant role in sup-optimum early pregnancy or mid-term abortion rates in deer.

Authors:Valeria Carolina Colombo; Ignacio Gamietea; Sylvia Grune Loffler; Bibiana Fel Brihuega; Pablo Martin BeldomenicoPages: 90 - 92Abstract: Publication date: February 2018 Source:Veterinary Microbiology, Volume 215 Author(s): Valeria Carolina Colombo, Ignacio Gamietea, Sylvia Grune Loffler, Bibiana Fel Brihuega, Pablo Martin Beldomenico We investigated the presence of infection by Leptospira spp. in an assembly of Sigmodontinae rodents from the Paraná Delta, Argentina. Rodents were captured in places with natural grassland, implanted forest, with and without raising cattle and in sites prone and not prone to flooding. The DNA was amplified from cultured isolates by PCR and Leptospira spp. strains were genotyped using Multiple – Locus Variable Number Tandem Repeat Analysis (MLVA). We isolated Leptospira interrogans serovar Copenhageni from Oligoryzomys nigripes, Leptospira borgpetersenii from Scapteromys aquaticus and Leptospira interrogans serovar Icterohaemorrhagiae from Akodon azarae. The zoonotic Leptospira isolated and genotyped from O. nigripes and S. aquaticus are the first reports from these species. The geographic range of these rodent species include, in addition to Argentina, the countries of Paraguay, Uruguay and Brazil, suggesting that these rodents might be involved in the transmission of spirochetes in other regions. Human and animal health care professionals should be alert to the potential occurrence of leptospirosis in areas where these rodent species are present.

Authors:Abbey J. Steckler; Maria X. Cardenas-Alvarez; Megan K. Townsend Ramsett; Neil Dyer; Teresa M. BergholzPages: 93 - 97Abstract: Publication date: February 2018 Source:Veterinary Microbiology, Volume 215 Author(s): Abbey J. Steckler, Maria X. Cardenas-Alvarez, Megan K. Townsend Ramsett, Neil Dyer, Teresa M. Bergholz Listeria monocytogenes infections are an important disease of ruminants worldwide, causing encephalitis, septicemia, and abortions. Ruminant listeriosis can also pose a food safety risk due to the potential for L. monocytogenes to enter the food supply via the farm environment. Data on the genetic diversity of L. monocytogenes from ruminant clinical cases in the United States is limited. Our goal was to assess the genetic diversity of clinical listeriosis isolates from ruminants in the Upper Great Plains states, a population not well-studied, and compare this population to isolates from ruminants in New York State. Multi-locus sequence typing (MLST) was used to classify and compare the genetic diversity of the isolates from the two regions. Loci sequences were compared to all known sequence types using the Pasteur Institute L. monocytogenes MLST database. Four novel sequence types (ST) were identified among the Upper Great Plains isolates, and four new STs were classified in the New York collection. Five STs were found to be common across the 2 geographical regions; ST 1, 7, 191, and 204. Strains of ST 7 were most frequently isolated (7/46 isolates). Strains of ST 91 were all associated with fetal infections from the Upper Great Plains. Our results demonstrate that while there are some subtypes commonly found between the two geographic regions, there are also subtypes distinct to each region.

Authors:Sarah van Alen; Ursula Kaspar; Evgeny A. Idelevich; Robin Köck; Karsten BeckerPages: 7 - 12Abstract: Publication date: February 2018 Source:Veterinary Microbiology, Volume 214 Author(s): Sarah van Alen, Ursula Kaspar, Evgeny A. Idelevich, Robin Köck, Karsten Becker Problem addressed Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA), particularly of the clonal complex (CC) 398, emerged as zoonotic pathogens predominantly among humans with direct or indirect livestock contact, but also in healthcare settings. The factors contributing to the success of LA-MRSA are only poorly understood. Objective During the past years, the use of heavy metal compounds as feed-supplements was found to influence the co-selection of LA-MRSA in pig herds. This study aimed to determine the prevalence of zinc resistance among MRSA CC398 isolated from patients of a German university hospital located in a pig farming-dense area. Methods and approach In comparison to concurrent healthcare-associated MRSA (HA-MRSA), LA-MRSA CC398 comprising isolates from their first appearance in 2000 to recent isolates from 2014 were included. Results Among MRSA CC398, the overall resistance rate towards zinc chloride was 57% compared to only 3% among concurrently isolated HA-MRSA. Zinc resistance correlated with the presence of the czrC gene in 100% of the MRSA CC398 and in 67% of the HA-MRSA. Conclusions The zinc resistance rate in MRSA CC398 significantly increased from 2009 to 2014 with a maximum in 2014. Alarmingly, zinc resistance has become a frequent phenotype of human LA-MRSA in Germany potentially facilitating co-selection of antibiotic resistance genes.

Authors:Olga Andrievskaia; Claude Turcotte; Gloria Berlie-Surujballi; Hannah Battaion; Dara LloydPages: 44 - 50Abstract: Publication date: February 2018 Source:Veterinary Microbiology, Volume 214 Author(s): Olga Andrievskaia, Claude Turcotte, Gloria Berlie-Surujballi, Hannah Battaion, Dara Lloyd Two internationally recognised and standardised genotyping methods, mycobacterial interspersed repetitive unit and variable number tandem repeat analysis (MIRU-VNTR) and spoligotyping, were applied to characterise genetic variations among 137 Mycobacterium bovis isolates recovered from Canadian domestic and wild animals during 1985–2015. Spoligotyping generated seven types that were discriminated further into12 MIRU-VNTR types. The discriminatory power indexes were estimated as 0.71 and 0.77 for spoligotyping and MIRU-VNTR typing approaches, respectively. In total, 6 prominent clusters of isolates were observed by the genotyping schemes. Four genotype clusters were exclusively observed in farmed animals. Three of these four clusters were affiliated with localised tuberculosis outbreaks, and each cluster corresponded to a single specific spoligotype (SB0140, SB0673, and SB1069) and a MIRU-VNTR profile. The fourth genotype cluster, with spoligotype SB0265 which segregated into two MIRU-VNTR types, was associated with bovine tuberculosis outbreaks in several farms across Canada during 1990–2002. Two genotype clusters of M. bovis stains were associated with wildlife reservoirs: a spoligotype SB0130 with 3 unique MIRU-VNTR profiles were observed in wood bison in Wood Buffalo National Park, and unique spoligotypes SB1070 and 1071 represented by four MIRU-VNTR profiles were recovered from cervidae species in and around the Riding Mountain National Park of Manitoba. Genotyping data confirmed M. bovis transmission between wildlife and livestock in Manitoba in 1990–2008. Overall, notwithstanding the low level of genetic diversity of Canadian M. bovis strains, the spoligotyping and MIRU-VNTR typing were useful tools in monitoring transmission of endemic strains and defining new introductions to Canada. The majority of genotypes were most likely introduced into domestic animals through live animal trade, and subsequently eliminated as a result of bovine tuberculosis outbreak investigation and eradication activities.

Authors:Ismail Boukahil; Charles J. CzuprynskiAbstract: Publication date: Available online 12 February 2018 Source:Veterinary Microbiology Author(s): Ismail Boukahil, Charles J. Czuprynski Mannheimia haemolytica and Pasteurella multocida are two bacterial species implicated in the bovine respiratory disease complex (BRDC) that is costly to the beef and dairy cattle industries. Both bacterial species are thought to occupy a similar niche as commensals in the upper respiratory tract. Many bacteria are thought to exist as biofilms in their hosts, perhaps in close proximity with other bacterial species. We previously showed that M. haemolytica forms biofilm on bovine respiratory epithelial cells in vitro. We are interested in the possibility that M. haemolytica and P. multocida co-exist as biofilms in the upper respiratory tract of cattle. In this study, we begin to explore this possibility by assessing the ability of M. haemolytica and P. multocida to form a biofilm on bovine respiratory epithelial cells in vitro. We found that M. haemolytica and P. multocida are separately able to form biofilms on bovine respiratory epithelial cells, but mutually inhibit one another when incubated together as a biofilm. Both the biofilm matrix (crystal violet stain) and bacterial numbers (CFU and PCR) were reduced when M. haemolytica and P. multocida were incubated together on fixed epithelial cells. This inhibition does not appear to result from a soluble factor, as neither conditioned medium nor separation of the two species by a transwell filter membrane reproduced the effect. We infer that when located in close proximity on the epithelial surface, M. haemolytica and P. multocida mutually regulate one another.

Authors:Miranda Prats-van der Ham; Juan Tatay-Dualde; Ángel Gómez-Martín; Juan Carlos Corrales; Antonio Contreras; Antonio Sánchez; Christian de la FeAbstract: Publication date: Available online 11 February 2018 Source:Veterinary Microbiology Author(s): Miranda Prats-van der Ham, Juan Tatay-Dualde, Ángel Gómez-Martín, Juan Carlos Corrales, Antonio Contreras, Antonio Sánchez, Christian de la Fe Mycoplasma capricolum subsp. capricolum (Mcc) is one of the causative agents of contagious agalactia, and antimicrobial treatment is the most commonly applied measure to treat outbreaks of this disease. Macrolides and lincosamides bind specifically to nucleotides at domains II and V of the 23S rRNA. Furthermore, rplD and rplV genes encode ribosomal proteins L4 and L22, which are also implicated in the macrolide binding site. The aim of this work was to study the relationship between mutations in these genes and the acquisition of macrolide and lincosamide resistance in Mcc. For this purpose, in vitro selected resistant mutants and field isolates were studied. This study demonstrates the appearance of DNA point mutations at the 23S rRNA encoding genes (A2058G, A2059G and A2062C) and rplV gene (Ala89Asp) in association to high minimum inhibitory concentration values. Hence, it proves the importance of alterations in 23S rRNA domain V and ribosomal protein L22 as molecular mechanisms responsible for the acquisition of macrolide and lincosamide resistance in both field isolates and in vitro selected mutants. Furthermore, these mutations enable us to provide an interpretative breakpoint of antimicrobial resistance for Mcc at MIC 0.8 µg/ml.

Authors:Xiumei Wang; Yao Zhu; Xin Hua; Fuguang Chen; Changzhen Wang; Yanhe Zhang; Siguo Liu; Wanjiang ZhangAbstract: Publication date: Available online 6 February 2018 Source:Veterinary Microbiology Author(s): Xiumei Wang, Yao Zhu, Xin Hua, Fuguang Chen, Changzhen Wang, Yanhe Zhang, Siguo Liu, Wanjiang Zhang The objective of this study was to investigate the prevalence of the cfr gene in Escherichia coli isolates from domestic animals in Northeast China and to characterize the cfr-containing plasmids. Between June 2015 and April 2016, 370 E. coli isolates were collected from pigs, chickens, and dairy cows in Northeast China. Among these, 111 were florfenicol resistant, including 109 isolates carrying the floR gene and 6 positives for cfr. The prevalence of cfr in E. coli isolates from the four northeast provinces in China was 1.6% (6/370), which was higher than that previously reported (0.08% and 0.5%). All six cfr-containing E. coli isolates were highly resistant to florfenicol (100%), cefotaxime (100%), and fosfomycin (100%). Complete sequence analysis of two cfr-carrying plasmids revealed high homology of the IncX4-type pEC14cfr plasmid with two other cfr-harboring plasmids, pSD11 and pGXEC6, found in swine E. coli isolates from southern China. pEC14cfr-like plasmids have been isolated in five provinces in southern and northern China. The isolation sites were up to 2700 kilometers apart, implying that pEC14cfr-like plasmids are likely to be national epidemic cfr-carrying plasmids that mediate the dissemination of cfr in China. Moreover, the genetic structure (IS26-IS26-cfr-rec-pre/mob-ramA-IS26) of the second cfr-carrying plasmid, IncF14:A-:B- pEC295cfr, represents a novel genetic environment for cfr identified for the first time in the present study. Sequence homology analysis indicated that the cfr-carrying element was most likely introduced into a cfr-negative pEC12 plasmid backbone, which evolved into the cfr-carrying vector, pEC295cfr. Moreover, isolation of the IncF14:A-:B- pEC295cfr plasmid harboring cfr suggests that IncFII plasmids maybe have become additional effective vehicles for cfr dissemination. These results highlight the importance of surveying the prevalence of IncX4 and IncFII plasmids in gram-negative bacteria, especially in swine E. coli isolates.

Authors:Silke Amelung; Maria Hartmann; Ludwig Haas; Lothar KreienbrockAbstract: Publication date: Available online 5 February 2018 Source:Veterinary Microbiology Author(s): Silke Amelung, Maria Hartmann, Ludwig Haas, Lothar Kreienbrock In Germany, all calves are tested for the presence of bovine viral diarrhoea/mucosal disease virus (BVDV) virus since January 1, 2011. The basis for this compulsory investigation is the BVDV Federal Regulation (BVDVV), which demands testing of calves before the age of six months and according to the new regulation of June 2016 within four weeks or before entering another stock. In 2012, a questionnaire was sent to 7,250 Lower Saxony cattle farmers to identify potential factors associated with the presence of BVDV. Completed questionnaires were received from 2,542 farms for further analysis. For BVD status determination of these farms, the diagnostic results of 425,911 ear notch samples of calves as part of the BVD eradication period from June 2010 to December 2013 were used. For the analysis of the completed questionnaires, a univariable analysis was performed by the chi-square or Wilcoxon test for each variable studied. In addition, a multivariable logistic model was performed. Four potential risk factors remained after a backward selection in the final logistic regression model: the dairy production compared to the suckling and other types of production, the herd size, the purchase of animals and the location in western region in comparison with the central and eastern regions. In summary, according to the results of this study, the farm with the highest probability of a BVDV infection in Lower Saxony is a large dairy farm that purchases cattle and is located in a cattle-dense region. When the complete eradication of the virus will be achieved, the results of the present study may help to conduct a risk-oriented monitoring programme.

Authors:Troels Ronco; Ilka C. Klaas; Marc Stegger; Line Svennesen; Lærke B. Astrup; Michael Farre; Karl PedersenAbstract: Publication date: Available online 9 January 2018 Source:Veterinary Microbiology Author(s): Troels Ronco, Ilka C. Klaas, Marc Stegger, Line Svennesen, Lærke B. Astrup, Michael Farre, Karl Pedersen Staphylococcus aureus is one of the most common pathogens that cause mastitis in dairy cows. Various subtypes, virulence genes and pathogenicity islands have been associated with isolates from bulk tank milk and clinical mastitis. So far, no Danish cattle associated S. aureus isolates have been whole-genome sequenced and further analyzed. Thus, the main objective was to investigate the population structure and genomic content of isolates from bulk tank milk and clinical mastitis, using whole-genome sequencing. This may reveal the origin of strains that cause clinical mastitis. S. aureus isolates from bulk tank milk (n = 94) and clinical mastitis (n = 63) were collected from 91 and 24 different farms, respectively and whole-genome sequenced. The genomic content was analyzed and a phylogenetic tree based on single nucleotide polymorphisms was constructed. In general, the isolates from both bulk tank milk and clinical mastitis were of similar genetic background. This suggests that dairy cows are natural carriers of the S. aureus subtypes that cause clinical mastitis if the right conditions are present and that a broad range of subtypes cause mastitis. A phylogenetic cluster that mostly consisted of ST151 isolates carried three pathogenicity islands that were primarily found in this group. The prevalence of resistance genes was generally low. However, the first ST398 methicillin resistant S. aureus isolate from a Danish dairy cow with clinical mastitis was detected.

Authors:Gagandeep Singh; Sheela RamamoorthyAbstract: Publication date: Available online 5 January 2018 Source:Veterinary Microbiology Author(s): Gagandeep Singh, Sheela Ramamoorthy Torque teno viruses [TTVs] are negative sense, single-stranded, DNA viruses, which are distributed globally in several mammalian hosts such as humans, apes, sheep and swine in a species-specific manner. While the pathogenic potential of TTVs is under debate, recent experimental studies in gnotobiotic pigs indicate that swine TTVs, TTSuV1 in particular, can act as a primary or co-infecting pathogen. Hence, determining whether TTSuV1 can infect other mammals would eventually further our understanding of viral pathogenesis, especially in coinfections. In this study, we tested sera from horses, cattle, sheep, dogs and elk for the presence of TTSuV1 DNA using a panel of TTSuV1-specific primers, and assessed the extent of sero-conversion to TTSuV1 in the selected species. We found that TTSuV1 DNA was detected in 46.7% of equines, 70% of canine, 100% of bovine, 40% of ovine and 93.3% of elk samples. However, significant TTSuV1 specific antibody responses were detected only in the bovine, ovine and equine samples but not the canine or elk samples, indicating that these animals could support the replication of TTSuV1. This combined serological and molecular epidemiological profile of TTSuV1 infection in five different species indicates the host range of species-specific TTVs could be wider than initially believed. Further studies are required to understand the health risks to these animal species from TTSuV-1 infection.

Authors:Aruna Amarasinghe; Mohamed Sarjoon Abdul-Cader; Zahraa Almatrouk; Frank van der Meer; Susan C. Cork; Susantha Gomis; Mohamed Faizal Abdul-CareemAbstract: Publication date: Available online 3 January 2018 Source:Veterinary Microbiology Author(s): Aruna Amarasinghe, Mohamed Sarjoon Abdul-Cader, Zahraa Almatrouk, Frank van der Meer, Susan C. Cork, Susantha Gomis, Mohamed Faizal Abdul-Careem Infectious bronchitis virus (IBV) infection is a major cause of economic losses for the poultry industry. Due to limitations in current control measures, alternative approaches, based on through understanding of the host responses are required. As one of the key component of the avian immune system, the innate immune system has a crucial role in limiting virus replication at the initial stage of the infection. As parts of the innate host response, macrophages and cytokines, such as interleukin (IL)-1β, are critical components as shown in other host-virus infection models. Since information on the importance of macrophages and IL-1β in IBV infection in chickens is limited, our objective was to determine the association of IL-1β, originating from avian macrophages and IBV infection in the trachea and lung. Following experimental IBV infection in 6 days old chickens, we found increased production of IL-1β and increased recruitment of macrophages in the respiratory tract. Towards the end of the study (5 and 7 days following the IBV infection), the recruited macrophages appear to be a significant source IL-1β. However, only the recruitment of macrophages in the lung correlated with IBV genome loads in this tissue. In conclusion, the present study demonstrates that recruitment of macrophages and the production of IL-1β originating from macrophages, as well as other sources, occur following IBV infection in the respiratory tract suggesting potential roles of these mediators in the host responses to IBV infection. However, further studies are warranted to elucidate whether macrophages and IL-1β are the causes of reduced IBV genome loads in the respiratory tract and also to investigate whether immune mediators that were not measured in the current study were involved in reducing IBV genome load in the respiratory tract towards the end of the study.

Authors:Anna Wanecka; Jarosław Król; Jan Twardoń; Jacek Mrowiec; Jacek Bania; Agnieszka Korzeniowska-Kowal; Anna TobiaszPages: 28 - 35Abstract: Publication date: February 2018 Source:Veterinary Microbiology, Volume 214 Author(s): Anna Wanecka, Jarosław Król, Jan Twardoń, Jacek Mrowiec, Jacek Bania, Agnieszka Korzeniowska-Kowal, Anna Tobiasz The aim of this paper is to describe a novel subpopulation of Staphylococcus haemolyticus isolated from intramammary gland infections (IMI) in cattle. In total, eight isolates originating from milk samples from two unrelated dairy farms were examined phenotypically (using the ID 32 STAPH system) and genotypically. These isolates had almost identical sequences of each of the housekeeping genes examined (dnaJ, rpoB and sodA) but these sequences displayed similarity of only ∼92.5%, 95.0% and 96.8%, respectively, with known S. haemolyticus sequences. The atypical isolates could also be distinguished biochemically by the positive β-galactosidase test (with 2-naphthyl-β-d-galactopyranoside as the substrate). All the isolates were identified as S. haemolyticus upon MALDI-TOF analysis but half of them, that achieved scores 1.7–1.999 (not reliable species identification), required expanding the commercial database for secure identification. Our study has shown that IMI in cattle may be caused by two distinct subpopulations of S. haemolyticus, differing clearly by some genotypic and phenotypic properties. The first of these subpopulations seems to be common to many hosts (including humans), whereas the second (possibly at the subspecies rank) is, so far, found only in cattle.

Authors:Anping Wang; Lingling Gu; Wu shuang; Shanyuan ZhuAbstract: Publication date: Available online 26 December 2017 Source:Veterinary Microbiology Author(s): Anping Wang, Lingling Gu, Wu shuang, Shanyuan Zhu Duck hepatitis A virus (DHAV), a non-enveloped ssRNA virus, can cause a highly contagious disease in young ducklings. The three capsid proteins of VP0, VP1 and VP3 are translated within a single large open reading frame (ORF) and hydrolyzed by protease 3CD. However, little is known on whether the recombinant viral structural proteins (VPs) expressed in insect cells could spontaneously assemble into virus-like particles (VLPs) and whether these VLPs could induce protective immunity in young ducklings. To address these issues, the structural polyprotein precursor gene P1 and the protease gene 3CD were amplified by PCR, and the recombinant proteins were expressed in insect cells using a baculovirus expression system for the characterization of their structures and immunogenicity. The recombinant proteins expressed in Sf9 cells were detected by indirect immunofluorescence assay and Western blot analysis. Electron microscopy showed that the recombinant proteins spontaneously assembled into VLPs in insect cells. Western blot analysis of the purified VLPs revealed that the VLPs were composed with the three structural proteins. In addition, vaccination with the VLPs induced high humoral immune response and provided strong protection. Therefore, our findings may provide a framework for development of new vaccines for the prevention of duck viral hepatitis.

Authors:Liu Jianchai; Liu Huanzhang; Yan Jinkun; Liu Na; Zhang Heping; Zhao Chengrui; Liu YanweiAbstract: Publication date: Available online 23 November 2017 Source:Veterinary Microbiology Author(s): Liu Jianchai, Liu Huanzhang, Yan Jinkun, Liu Na, Zhang Heping, Zhao Chengrui, Liu Yanwei Candida albicans is the most prevalent opportunistic fungus of humans and animals. While most studies focus on human isolates, they rarely focus on poultry isolates. In this study, C. albicans strains were recovered from poultry in the southern Hebei Province (China) and identified. Molecular typing and analyses were performed to understand the molecular epidemiology and genetic relatedness of the strains. The fungi were isolated from live birds with presumed candidiasis or their corpses. The isolates were identified based on morphology, differential medium culture, and rDNA internal transcribed spacer sequencing. The identified C. albicans strains were analyzed by ABC genotyping and multilocus sequence typing. Clonal groups were identified using the eBURST (version 3.0) software, and an UPGMA phylogenetic tree was constructed using the MEGA (version 6.06) software. Overall, 72 isolates were divided into three genotypes (A, B, and C), 48 novel sequence types (STs), five groups with 10 singletons, and four clades. Results indicated that candidiasis is common in poultry in the southern Hebei Province, and that the genetic composition of the C. albicans poultry population from the area is relatively complicated. Based on the eBURST analysis for the STs in this study and others, we suggest that C. albicans poultry isolates were relatively independent but not completely separated from human isolates. The strains with the same or closely related genotypes but recovered from both birds and humans could have transferred and evolved between the two types of host.